Phenotypic characterization of DC subsets in secondary lymphoid organs.

<p>Single cell suspensions from retro-pharyngeal, sub-mandibular, pre-scapular, popliteal lymph nodes, and a spleen were prepared, and 7-color flow cytometric analysis performed to identify DC. Doublets, dead cells, and lineage positive cells (T cells, monocytes, B cells and NK cells) were excluded as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0109273#pone-0109273-g001" target="_blank">Figure 1</a>. Lineage negative cells were then gated to identify MHC class II⁺ and CD4⁺ cells (A), and the MHC class II⁺ CD4⁻ cells were assessed for CD11c expression (B). Surface expression of DEC205 and CD172a by CD4⁺ MHC class II⁻ DC (C), MHC class II⁺/CD4⁻/CD11c⁺ DC (D), and MHC class II⁺/CD4⁻/CD11c⁻ DC (E) were assessed. Numbers on plots represent percentage of cells expressing the surface markers shown.